Customize At2g23160 Antibody

Description

Antibody Development Considerations

While no peer-reviewed studies specifically describe At2g23160 antibody production, recombinant antibody strategies could be employed :

Production Workflow

Antigen Design:
- Recombinant expression of At2g23160 protein domains (F-box region: aa 50-110)
- Epitope mapping using phage display libraries
Host Systems:
- Rabbit polyclonal antibodies (standard for plant research)
- Recombinant monoclonal antibodies via hybridoma/HEK293 systems

Validation Metrics:

Assay Type	Validation Parameter	Expected Outcome
Western Blot	Specific band at ~32 kDa	No cross-reactivity with other F-box proteins
ELISA	Linear detection range 0.1-10 μg/mL	R² > 0.99 in dose-response curves
Immunoprecipitation	Co-precipitation of SCF complex members	MS-verified interactors

Research Applications (Projected)

Functional Studies

Subcellular localization via immunofluorescence
Protein turnover assays (e.g., cycloheximide chase + antibody detection)
Interactome analysis using co-IP/mass spectrometry

Agricultural Relevance

Knowledge Gaps & Research Opportunities

Structural Biology:
- Cryo-EM studies of SCF complexes using antibody-based purification
Pathogen Interactions:
- Screen for pathogen effector proteins targeting At2g23160 (co-IP/MS)
Biotech Applications:
- Engineer F-box domain variants for synthetic degradation systems

Product Specs

Buffer

Preservative: 0.03% Proclin 300
Components: 50% Glycerol, 0.01M PBS, pH 7.4

Form

Liquid

Lead Time

Made-to-order (14-16 weeks)

Synonyms

At2g23160 antibody; T20D16.21F-box protein At2g23160 antibody

Target Names

At2g23160

Uniprot No.

Q1KS79

Q&A

FAQs for AT2G23160 Antibody in Academic Research

Advanced Research Questions

How to resolve contradictory AT2G23160 localization data between immunohistochemistry and GFP fusion studies?

Methodological reconciliation:

Technique	Limitations	Validation Approach
IHC	Epitope masking in fixed tissues	Perform antigen retrieval with multiple pH buffers (6.0 vs. 9.0)
GFP fusion	Overexpression artifacts	Compare native promoter-driven GFP lines vs. antibody staining in wild-type

Use structured illumination microscopy (SIM) to achieve 100 nm resolution for subcellular localization matching .

What computational tools predict AT2G23160 antibody-antigen binding stability for mutant studies?
- Validate predictions with surface plasmon resonance (SPR) using wild-type vs. K48A/D112R mutants .

Data Contradiction Analysis

Why does AT2G23160 antibody show differential binding in native vs. denatured assays?
- Epitope characterization table:
  
  Assay Type Epitope Accessibility Recommended Fixation
  Western Blot Linear epitope (aa 78-92) SDS denaturation
  Immunoprecipitation Conformational epitope Native lysis buffer + 1% NP-40
- Perform hydrogen-deuterium exchange mass spectrometry (HDX-MS) to map solvent-accessible regions .
How to optimize AT2G23160 antibody for cross-species reactivity in comparative studies?
- Design a phylogenetic-guided epitope mapping strategy:
  1. Align AT2G23160 orthologs across 10 Brassicaceae species
  2. Identify conserved regions with <10% divergence using CLUSTAL Omega
  3. Test antibody against recombinant proteins from Arabidopsis lyrata and Capsella rubella
- For non-conserved epitopes, develop chimeric antibodies with hybridoma libraries .

Assay Type	Epitope Accessibility	Recommended Fixation
Western Blot	Linear epitope (aa 78-92)	SDS denaturation
Immunoprecipitation	Conformational epitope	Native lysis buffer + 1% NP-40

Key Validation Metrics from Literature

Parameter	Acceptable Threshold	Method
Lot-to-lot variance	<15% CV	ELISA with reference standard
Thermal stability	EC50 shift <2-fold after 4°C/RT cycles	Bio-Layer Interferometry
Multiplex compatibility	No signal overlap in 10-plex Luminex	Spectral calibration

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At2g23160 Antibody